LOCUS       sequence-84560-e        4190 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4190)
  AUTHORS   Binder A, Lambert J, Morbitzer R, Popp C, Ott T, Lahaye T, Parniske 
            M
  TITLE     A modular plasmid assembly kit for multigene expression, gene 
            silencing and silencing rescue in plants.
  JOURNAL   PLoS One. 2014 Feb 13;9(2):e88218. doi: 
            10.1371/journal.pone.0088218. eCollection 2014.
  PUBMED    24551083
REFERENCE   2  (bases 1 to 4190)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS One.";
            date: "2014-02-13"; volume: "9(2):e88218. doi"; pages: " 
            10.1371/journal.pone.0088218. eCollection 2014"
COMMENT     Sequence Label: LI+Bpi
FEATURES             Location/Qualifiers
     source          1..4190
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     466..484
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(522..544)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     644..663
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     857..879
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     871..888
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     872..888
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    967..1091
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(1116..1121,1122..1139,1140..1146)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             1200..1859
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1266..1285)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             2201..2506
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     2385..2404
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(2547..2671)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2714..2730)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2714..2730)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2727..2749)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2738..2754
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(2762..2768,2769..2786,2787..2792))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2807..2828
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2945..2962)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3116..3704)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3196..3215)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(3819..3824,3825..3841,3842..3847)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             join(4036..4190,1..379)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
ORIGIN
        1 cgacgtagcc acctactccc aacatcagcc ggactccgat tacctcggga acttgctccg
       61 tagtaagaca ttcatcgcgc ttgctgcctt cgaccaagaa gcggttgttg gcgctctcgc
      121 ggcttacgtt ctgcccaggt ttgagcagcc gcgtagtgag atctatatct atgatctcgc
      181 agtctccggc gagcaccgga ggcagggcat tgccaccgcg ctcatcaatc tcctcaagca
      241 tgaggccaac gcgcttggtg cttatgtgat ctacgtgcaa gcagattacg gtgacgatcc
      301 cgcagtggct ctctatacaa agttgggcat acgggaagaa gtgatgcact ttgatatcga
      361 cccaagtacc gccacctaac aattcgttca agccgagatc ggcttcccgg ccgcggagtt
      421 gttcggtaaa ttgtcacaac gccgctcatg acattaacct ataaaaatag gcgtatcacg
      481 aggccctttt gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc
      541 ccggagaagg tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc
      601 gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt
      661 gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac
      721 cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg
      781 gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg
      841 gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga attcgagctc
      901 ggtacctcgc gaatgcatct agatatcgga tcccgggccc gtcgactgca gaggtacgaa
      961 gtcttcacaa gtttgtacaa aaaagctgaa cgagaaacgt aaaatgatat aaatatcaat
     1021 atattaaatt agattttgca taaaaaacag actacataat actgtaaaac acaacatatc
     1081 cagtcactat ggcggccgca ttaggcaccc caggctttac actttatgct tccggctcgt
     1141 ataatgtgtg gattttgagt taggatccgt cgagattttc aggagctaag gaagctaaaa
     1201 tggagaaaaa aatcactgga tataccaccg ttgatatatc ccaatggcat cgtaaagaac
     1261 attttgaggc atttcagtca gttgctcaat gtacctataa ccagaccgtt cagctggata
     1321 ttacggcctt tttaaagacc gtaaagaaaa ataagcacaa gttttatccg gcctttattc
     1381 acattcttgc ccgcctgatg aatgctcatc cggaattccg tatggcaatg aaagacggtg
     1441 agctggtgat atgggatagt gttcaccctt gttacaccgt tttccatgag caaactgaaa
     1501 cgttttcatc gctctggagt gaataccacg acgatttccg gcagtttcta cacatatatt
     1561 cgcaagatgt ggcgtgttac ggtgaaaacc tggcctattt ccctaaaggg tttattgaga
     1621 atatgttttt cgtctcagcc aatccctggg tgagtttcac cagttttgat ttaaacgtgg
     1681 ccaatatgga caacttcttc gcccccgttt tcaccatggg caaatattat acgcaaggcg
     1741 acaaggtgct gatgccgctg gcgattcagg ttcatcatgc cgtttgtgat ggcttccatg
     1801 tcggcagaat gcttaatgaa ttacaacagt actgcgatga gtggcagggc ggggcgtaaa
     1861 gatctggatc cggcttacta aaagccagat aacagtatgc gtatttgcgc gctgattttt
     1921 gcggtataag aatatatact gatatgtata cccgaagtat gtcaaaaaga ggtatgctat
     1981 gaagcagcgt attacagtga cagttgacag cgacagctat cagttgctca aggcatatat
     2041 gatgtcaata tctccggtct ggtaagcaca accatgcaga atgaagcccg tcgtctgcgt
     2101 gccgaacgct ggaaagcgga aaatcaggaa gggatggctg aggtcgcccg gtttattgaa
     2161 atgaacggct cttttgctga cgagaacagg ggctggtgaa atgcagttta aggtttacac
     2221 ctataaaaga gagagccgtt atcgtctgtt tgtggatgta cagagtgata ttattgacac
     2281 gcccgggcga cggatggtga tccccctggc cagtgcacgt ctgctgtcag ataaagtctc
     2341 ccgtgaactt tacccggtgg tgcatatcgg ggatgaaagc tggcgcatga tgaccaccga
     2401 tatggccagt gtgccggtct ccgttatcgg ggaagaagtg gctgatctca gccaccgcga
     2461 aaatgacatc aaaaacgcca ttaacctgat gttctgggga atataaatgt caggctccct
     2521 tatacacagc cagtctgcag gtcgaccata gtgactggat atgttgtgtt ttacagtatt
     2581 atgtagtctg ttttttatgc aaaatctaat ttaatatatt gatatttata tcattttacg
     2641 tttctcgttc agctttcttg tacaaagtgg ttgggaagac attctgcctg catgcaagct
     2701 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     2761 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     2821 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     2881 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     2941 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     3001 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     3061 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     3121 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     3181 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     3241 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     3301 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     3361 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     3421 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     3481 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     3541 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     3601 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     3661 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     3721 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     3781 ggcgcacacc gtggaaacgg atgaaggcac gaacccagtt gacataagcc tgttcggttc
     3841 gtaaactgta atgcaagtag cgtatgcgct cacgcaactg gtccagaacc ttgaccgaac
     3901 gcagcggtgg taacggcgca gtggcggttt tcatggcttg ttatgactgt ttttttgtac
     3961 agtctatgcc tcgggcatcc aagcagcaag cgcgttacgc cgtgggtcga tgtttgatgt
     4021 tatggagcag caacgatgtt acgcagcagc aacgatgtta cgcagcaggg cagtcgccct
     4081 aaaacaaagt taggtggctc aagtatgggc atcattcgca catgtaggct cggccctgac
     4141 caagtcaaat ccatgcgggc tgctcttgat cttttcggtc gtgagttcgg
//
